Microbial-induced carbon competition in the spermosphere leads to pathogen and disease suppression in a municipal biosolids compost

The aim of this study was to understand whether competition for fatty acids in plant seed exudates by compost-derived seed-colonizing microbial communities could explain the suppression of plant infections initiated by sporangia of Pythium ultimum. The germination behavior of P. ultimum sporangia in response to cucumber seeds was measured to determine the impact of seed-colonizing microbes on pathogen suppression. Seed-colonizing microbial communities from municipal biosolids compost utilized cucumber seed exudates and linoleic acid in vitro, reducing the respective stimulatory activity of these elicitors to P. ultimum sporangial germination. However, when sporangia were observed directly in the spermosphere of seeds sown in the compost medium, levels of germination and sporangial emptying did not differ from the responses in sand. The percentage of aborted germ tubes was greater after incubating sporangia in compost medium for 12-h than the level of germ tube abortion when sporangia were incubated in sand. Abortion did not occur if previously germinated sporangia were supplemented with cucumber seed exudate. Furthermore, removal of cucumber seed exudate after various stages of germ tube emergence resulted in an increase in aborted germ tubes over time. Adding increasing levels of glucose directly to the compost medium alleviated germ tube abortion in the spermosphere and also eliminated disease suppression. These data fail to support a role for linoleic acid competition in Pythium seedling disease suppression but provide evidence for general carbon competition mediated by seed-colonizing microbial communities as a mechanism for the suppression of Pythium seed infections in municipal biosolids compost.     

阴沟肠杆菌MY01的鉴定及防治水稻细菌性条斑病研究

 肠杆菌属(Enterobacter sp.)作为有益微生物的重要菌种,不仅对一些植物的生长起着促进作用,而且在有害生物防治方面起着重要的作用。本研究采用Biolog微生物生理生化分析、共培养等手段,对阴沟肠杆菌MY01、抗生素溶杆菌13-6与水稻细菌性条斑病菌RS105的营养和空间竞争能力、定殖特性以及对水稻幼苗的促生作用、条斑病的生防效果进行测定,明确阴沟肠杆菌与抗生素溶杆菌对水稻条斑病的协同生防作用。结果显示MY01生长迅速且能利用大部分碳源,营养竞争能力较强,可限制其他细菌生长。温室喷雾接种时,MY01与13-6对水稻条斑病的防效相近,且MY01可增强13-6对水稻条斑病的相对防效,防效高达93.44%。阴沟肠杆菌MY01作为一种潜在的生防菌株,对水稻条斑病具有一定防效,且促进抗生素溶杆菌13-6对水稻条斑病的防治。     

Interactions Between Trichoderma harzianum Strain T22 and Maize Inbred Line Mo17 and Effects of These Interactions on Diseases Caused by Pythium ultimum and Colletotrichum graminicola

ABSTRACT Seed treatment with Trichoderma harzianum strain T22, which results in colonization of plant roots but little or no colonization of shoots or leaves, had substantial effects on growth of and disease expression in maize inbred line Mo17. Shoots and roots of 10-day-old seedlings grown in a sandy loam field soil were larger (roots were nearly twice as long) in the presence of T22 than in its absence. Both main and secondary roots were increased in size and area and the root hair area was greater with T22. However, root hair area per unit of root length was greater in control plants. Increased growth probably was due to direct stimulation of plant growth in addition to effects from biological control of deleterious microflora. Seedlings of Mo17 grown in autoclaved or mefenoxamtreated sandy loam field soil were larger than those produced in untreated soil. However, seedlings grown in the presence of T22, either in treated or untreated soil, were larger than those produced in its absence. Infestation of soil with Pythium ultimum had little effect upon growth of Mo17. The presence of T22 increased protein levels and activities of beta-1,3 glucanase, exochitinase, and endochitinase in both roots and shoots, even though T22 colonized roots well but colonized shoots hardly at all. With some enzymes, the combination of T22 plus P. ultimum gave the greatest activity. Plants grown from T22-treated seed had reduced symptoms of anthracnose following inoculation of leaves with Colletotrichum graminicola, which indicates that root colonization by T22 induces systemic resistance in maize.     

Evaluation of the Endophyte Enterobacter cloacae SM10 Isolated from Spinach Roots for Biological Control against Fusarium Wilt of Spinach

Six hundred sixty-three isolates of microorganisms, including fungi and bacteria, were collected from surface-sterilized roots of spinach (Spinacia oleracea L.) growing in commercial greenhouses in Kyoto Prefecture. These isolates were screened for their ability to control Fusarium wilt of spinach caused by Fusarium oxysporum f. sp. spinaciae. In primary screening, spinach seeds were treated with the isolates, sown in pots containing sterilized soil, and then challenge-inoculated with the pathogen. Nine bacteria were effective in reducing disease incidence. Subsequently, spinach seeds were treated with the selected isolates, then sown in an infested field and grown from June to July 1998. Four bacteria reduced disease incidence. One of these four, designated as SM10, significantly suppressed the disease. Based on bacteriological properties, SM10 was identified as a strain of Enterobacter cloacae. SM10 was observed within xylem vessels of spinach roots using light and immunoelectron microscopy, indicating E. cloacae SM10 was an endophytic bacterium of spinach. Received 4 July 2000/ Accepted in revised form 13 September 2000     

Biotic Factors Affecting Expression of the 2,4-Diacetylphloroglucinol Biosynthesis Gene phlA in Pseudomonas fluorescens Biocontrol Strain CHA0 in the Rhizosphere

ABSTRACT Production of the polyketide antimicrobial metabolite 2,4-diacetyl-phloroglucinol (DAPG) is a key factor in the biocontrol activity of Pseudomonas fluorescens CHA0. Strain CHA0 carrying a translational phlA'-'lacZ fusion was used to monitor expression of the phl biosynthetic genes in vitro and in the rhizosphere. Expression of the reporter gene accurately reflected actual production of DAPG in vitro and in planta as determined by direct extraction of the antimicrobial compound. In a gnotobiotic system containing a clay and sand-based artificial soil, reporter gene expression was significantly greater in the rhizospheres of two monocots (maize and wheat) compared with gene expression in the rhizospheres of two dicots (bean and cucumber). We observed this host genotype effect on bacterial gene expression also at the level of cultivars. Significant differences were found among six additional maize cultivars tested under gnotobiotic conditions. There was no difference between transgenic maize expressing the Bacillus thuringiensis insecticidal gene cry1Ab and the near-isogenic parent line. Plant age had a significant impact on gene expression. Using maize as a model, expression of the phlA'-'lacZ reporter gene peaked at 24 h after planting of pregerminated seedlings, and dropped to a fourth of that value within 48 h, remaining at that level throughout 22 days of plant growth. Root infection by Pythium ultimum stimulated bacterial gene expression on both cucumber and maize, and this was independent of differences in rhizosphere colonization on these host plants. To our knowledge, this is the first comprehensive evaluation of how biotic factors that commonly confront bacterial inoculants in agricultural systems (host genotype, host age, and pathogen infection) modulate the expression of key biocontrol genes for disease suppression.     

Colonization of Pythium oligandrum in the tomato rhizosphere for biological control of bacterial wilt disease analyzed by real-time PCR and confocal laser-scanning microscopy

It recently has been reported that the non-plant-pathogenic oomycete Pythium oligandrum suppresses bacterial wilt caused by Ralstonia solanacearum in tomato. As one approach to determine disease-suppressive mechanisms of action, we analyzed the colonization of P. oligandrum in rhizospheres of tomato using real-time polymerase chain reaction (PCR) and confocal laser-scanning microscopy. The real-time PCR specifically quantified P. oligandrum in the tomato rhizosphere that is reliable over a range of 0.1 pg to 1 ng of P. oligandrum DNA from 25 mg dry weight of soil. Rhizosphere populations of P. oligandrum from tomato grown for 3 weeks in both unsterilized and sterilized field soils similarly increased with the initial application of at least 5 x 10(5) oospores per plant. Confocal microscopic observation also showed that hyphal development was frequent on the root surface and some hyphae penetrated into root epidermis. However, rhizosphere population dynamics after transplanting into sterilized soil showed that the P. oligandrum population decreased with time after transplanting, particularly at the root tips, indicating that this biocontrol fungus is rhizosphere competent but does not actively spread along roots. Protection over the long term from root-infecting pathogens does not seem to involve direct competition. However, sparse rhizosphere colonization of P. oligandrum reduced the bacterial wilt as well as more extensive colonization, which did not reduce the rhizosphere population of R. solanacearum. These results suggest that competition for infection sites and nutrients in rhizosphere is not the primary biocontrol mechanism of bacterial wilt by P. oligandrum.     

Influence of plant species on disease suppression by Pseudomonas fluorescens strain CHAO with enhanced antibiotic production

Introduction of the recombinant cosmid pME3090 into Pseudomonas fluorescens strain CHAO, a good biocontrol agent of various diseases caused by soilborne pathogens, increased three- to five-fold the production of the antibiotic metabolites pyoluteorin (Pit) and 2,4-diacetylphlorogIucinol (Phi) in vitro. Strain CHAO/pME3090 also overproduced Pit and Phi in the rhizosphere of wheat infected or not infected with Pythium ultimum. The biocontrol activity of the wild-type and recombinant Straitis was compared using various plant pathogen-host combinations in a gnotobiotic system. Antibiotic overproduction affected neither the protection of wheat against P. ultimum and Gaeumannomyces graminis var. tritici nor the growth of wheat plants. In contrast, strain CHA0/pME3090 showed an increased capacity to protect cucumber against Fusarium oxysporum f. sp. cucumerinum and Phomopsis sclerotioides , compared with the wild-type strain CHAO, The antibiotic overproducing strain protected tobacco roots significantly better against Thielaviopsis basicola than the wild-type strain but drastically reduced the growth of tobacco plants and was also toxic to the growth of sweet com. On King's B agar and on malt agar, the recombinant strain CHA0/pME3090 inhibited all pathogens more than did the parental strain CHAO. Synthetic Pit and Phi were toxic to all fungi tested. Tobacco and sweet com were more sensitive to synthetic Pit and Phi than were cucumber and wheat. There was no correlation between the sensitivity of the pathogens to the synthetic antibiotics and the degree of disease suppression by strain CHAO pME3090. However, there was a correlation between the sensitivity of the plants and the toxicity of the recombinant strain. We conclude that the plant species rather than the pathogen determines whether cosmid pME3090 in P. fluorescens strain CHAO leads to improved disease suppression.     

Suppression of Seedling Damping-Off Caused by Pythium ultimum, P. irregulare, and Rhizoctonia solani in Container Media Amended with a Diverse Range of Pacific Northwest Compost Sources

ABSTRACT Suppression of seedling damping-off disease caused by Pythium spp. and Rhizoctonia solani is a potential benefit of formulating soilless container media with compost. Thirty-six compost samples from Pacific Northwest commercial composting facilities were analyzed for a number of physical, chemical, and biological properties, including suppression of damping-off caused by Pythium ultimum, P. irregulare, and R. solani. The samples were produced from diverse feedstocks and composting technol ogies; this was reflected in a large degree of variability in the measured properties. When mixed with sphagnum peat moss and inorganic aggregates, 67% of the compost samples significantly suppressed P. irregulare damping-off of cucumber, 64% suppressed P. ultimum damping-off of cucumber, and 17% suppressed damping-off of cabbage caused by R. solani. Suppression of Pythium damping-off was related to the potential of compost to support microbial activity and a qualitative index of ammonia volatilization. Suppression of Rhizoctonia damping-off was not related to any one compost factor. Currently available compost products potentially could provide commercially acceptable control of damping-off caused by Pythium spp., but it is necessary to fortify composts with microbial antagonists for the control of R. solani.     

生防菌EN5的定殖能力及其对根际土壤微生物类群的影响

［目的］ 明确生防菌EN5菌株在番茄根、茎及根际土壤中和在黄瓜、烟草体内的定殖情况,及对根际土壤微生态的影响,为细菌性青枯病的生态治理提供理论依据。［方法］ 采用抗生素抗性标记法测定生防芽胞杆菌EN5的定殖能力;以平板培养及ERIC PCR法分析菌株EN5对番茄根际土壤微生物种群的影响。［结果］ 菌株EN5在番茄根、茎及根际土壤中均能稳定定殖,当菌株EN5处理植株15 d时,其在番茄茎内的定殖量可达5.6×104 cfu/g;此外,菌株EN5在烟草和黄瓜体内亦可定殖,其在烟草体内的定殖量较大。菌株EN5处理使根际土壤中细菌、真菌和放线菌的数量明显高于对照土壤,对氨化细菌、固氮菌、纤维素降解菌等微生物群体有促进作用,对反硫化细菌群体有抑制作用。同时,菌株EN5处理还改善了根际土壤中细菌群体的多样性。［结论］ 菌株EN5可以在其自然寄主番茄的根、茎及根际土壤中稳定定殖。同时,菌株EN5处理有助于改善根际土壤微生态环境,抑制病菌繁殖。     

Pathogenicity of four Pythium species to wheat, barley, peas and lentils

Pythium heterothallicum, P. irregulare, P. torulosum and P. ultimum var. sporangiiferum were compared for pathogenicity to seedlings of winter wheat, spring barley, lentils and peas in growth chambers at 5, 10, 15, 20 and 25 C. These four fungi are among the most commonly isolated Pythium species from wheat roots and wheat-field soils in eastern Washington and northern Idaho, USA, where wheat, spring barley, lentils and peas are grown in various rotations. Pathogenicity was determined in artificially infested soils (500 propagules per g) based on ability to cause pre-emergence death and post-emergence stunting of seedlings. P. ultimum var. sporangiiferum caused significant pre-emergence death of the wheat at 15–25 C, lentils at 10–25 C. and peas at 5 25 C. P. irregulare caused pre-emergence death only of peas and only at 5 C. With the possible exception of lentils at 25 C, P. heterothallcum and P. torulosum caused no pre-emergence death of any of the four plant species. None of the species caused pre-emergence death of spring barley. P. ultimum var. sporangiiferum caused the most post-emergence stunting of wheat, peas and lentils at 10 C and above. Pythium irregulare caused as much or more stunting than P. ultimum var. sporangiiferum on wheat, lentils and peas at 5 C, and was the most pathogenic species on barley at 10, 20 and 25 C. P. irregulare caused significantly more post-emergence stunting of wheat at 5 C with than without chaff (added as a food base for the pathogen); this was not offset by adding ammonium sulphate with the chaff.     

Characteristics and pathogenicity of Pythium spp. associated with root rot of tomatoes in soilless culture in Brittany, France

Pythium species cause necrotic lesions and root rots on tomato in soilless culture. Epidemiological studies conducted in Brittany, France, revealed the precocity and the frequency of the contamination of tomato roots by these fungi, even in symptomless roots. The colonization, estimated by culture-plate methods, could reach levels as high as 10⁵ propagules/gram of root. The frequent occurrence of Pythium with filamentous non-inflated sporangia, representing 75% of the total isolates, was observed during both latent infections and necrotic phases. Those Pythium spp., grouped as Pythium F, induced variable necrotic lesions in tomato roots in soilless culture. This group, which had been overlooked previously in favour of known pathogenic species such as P. ultitnum var. ultimum , can be pathogenic to tomato plants and therefore form a component of a potential disease complex.     

普城沙雷菌A21-4根际定殖能力及其对辣椒生长和诱导抗病性的影响

 本文探讨了辣椒疫病生防菌普城沙雷菌A21-4在辣椒根际的定殖能力及其对辣椒的促生长和诱导抗病性影响。以10⁸cfu·mL^-1A21-4菌液处理辣椒苗,A21-4能够有效地定殖在辣椒根际土壤和辣椒根部,移栽第30 d在根际土壤和辣椒根部均保持10⁶cfu·g^-1以上定殖密度。辣椒根际土壤中A21-4的定殖密度和病原菌存在与否无显著差异,而辣椒根部A21-4的定殖密度在病原菌存在时显著高于没有病原菌的;A21-4处理有效促进了辣椒地上部和根部的各项生育指标,同时,显著提高了辣椒叶绿素含量和根系活力。A21-4处理的辣椒苗叶绿素含量和根系活力比对照各提高86.1%和481.8%;经A21-4处理后,辣椒根部和叶部的SOD、POD和PAL活性明显提高,辣椒根部的SOD、POD和PAL活性峰值分别比对照提高44.7%、64.2%和77.0%,辣椒叶部SOD、POD和PAL活性峰值分别比对照提高27.9%、134.9%和87.0%;此外,A21-4浸根处理还能够提高辣椒叶部对辣椒疫霉菌的抗性。     

Population Dynamics of Bacillus sp. L324-92R(12) and Pseudomonas fluorescens 2-79RN(10) in the Rhizosphere of Wheat

ABSTRACT Bacillus sp. L324-92 is suppressive to three root diseases of wheat, namely take-all caused by Gaeumannomyces graminis var. tritici, Rhizoctonia root rot caused by Rhizoctonia solani AG8, and Pythium root rot caused by several Pythium species. Populations of strain L324-92R(12), a rifampicin-resistant mutant of L324-92 applied as a seed treatment, were monitored in the rhizosphere and spermosphere of wheat and compared with populations of Pseudomonas fluorescens 2-79RN(10), a known, rhizosphere-competent, biocontrol agent. In growth chamber studies, the population sizes of L324-92R(12) on roots of wheat were approximately 1,000-fold smaller than those of 2-79RN(10) at 5 days after planting, but, thereafter, they increased while those of 2-79RN(10) decreased until the two were equal in size at 45 days after planting. In the field with winter wheat, the population sizes of L324-92R(12) on roots were at least 10-fold smaller than those of 2-79RN(10) during the fall (November 1993) and early spring (March 1994). Thereafter, the population of L324-92R(12) remained constant or increased slightly, while the population of 2-79RN(10) decreased until the two were roughly the same at 10(4) to 10(5) CFU/plant over the period of 150 days (April 1994) until 285 days (harvest) after planting. In growth chamber studies, strain L324-92R(12) remained confined to root sections within 3.5 cm below the seed, whereas 2-79RN(10) was recovered from all root sections ranging from 0.5 to 6.5 cm below the seed. In the field on winter wheat, both strains were recovered from root sections down to 5.0 to 6.5 cm below the seed at 75 days after planting (mid December), but only 2-79RN(10) was recovered at this depth at 90 days after planting. Both strains were recovered from the seed remnants 6 months after planting in the field. Both strains also were recovered from inside the roots and shoots, but population sizes of strain 279RN(10) were greater than those of L324 92R(12).     

GFP标记生防细菌B579及其定殖能力检测

 利用绿色荧光蛋白（Green fluorescent protein, GFP）标记靶标微生物是目前研究微生物和宿主互作的重要手段。在本研究中,作者用电击转化的方法将携带质粒gfpmut3a基因的穿梭载体pGFP4412导入生防枯草芽孢杆菌（Bacillus subtilis）B579中,并得到成功表达GFP的枯草芽孢杆菌B579-gfp。用抗生素平板回收结合激光扫描共聚焦显微镜（LSCM）观察对枯草芽孢杆菌B579-gfp在盆栽的黄瓜根表定殖情况进行了研究,结果表明：标记菌株在激发光波长为488 nm的蓝光下可观察到亮绿色的荧光;B579-gfp菌体能够定殖在黄瓜根部,在根基部和中部都有菌体聚集而形成膜状结构,在根的分叉和根冠处可观察到大量B579-gfp定殖;浸种、蘸根以及灌根接种均可回收到大量的B579-gfp,分别为4.0×10³、1.0×10⁴和2.0×10² cfu/g。     

Endo-1,3-β-glucanase and cellulase from Trichoderma harzianum: purification and partial characterization, induction of and biological activity against plant pathogenic Pythium spp.

There were indications that endo-1,3--glucanase (1,3-(1,3;1,4)--D-Glucan 3(4)-glucanohydrolase (EC 3.2.1.6)) and cellulase (1,4-(1,3;1,4)--D-Glucan 4-glucanohydrolase (EC 3.2.1.4)) activity of Trichoderma harzianum Rifai isolate T3 were induced in sphagnum peat moss cultivations and dual culture experiments by the presence of Pythium ultimum. Further, P. ultimum stimulated the germination of Trichoderma conidia. Endo-1,3--glucanase and cellulase were purified from T. harzianum isolate T3, known to control Pythium damping-off of cucumber seedlings. The enzymes were purified from the culture filtrate of the fungus by gel filtration and isoelectric focusing. The purified endo-1,3--glucanase was a small protein with a molecular mass of 17 kilodaltons and a pI of 5.0. Two cellulases were purified to homogeneity and had molecular masses of 40 and 45 kilodaltons respectively, and pI's of 6.4 and 7.6 respectively. Germination of encysted zoospores and elongation of germ tubes of a plant pathogenic Pythium isolate were inhibited by low concentrations of the purified enzymes. A strong synergistic effect was observed on the inhibition of cyst germination by a combination of the endo-1,3--glucanase and the fungicide Fongarid. Finally, a time-course study of colonization of the rhizosphere of cucumber seedlings showed that the active fungal mycelial biomass of a GUS-transformant of T. harzianum isolate T3 increased over four weeks. Trichoderma appeared to colonize healthy roots only superficially, whereas the mucilage of the root hairs and of distal parts of wounded areas or broken parts of the roots, were extensively colonized.     

枯草芽胞杆菌PTS-394的GFP标记及其定殖能力

为探明枯草芽胞杆菌PTS-394在番茄根围的定殖能力,采用电转化法获得绿色荧光蛋白（green fluorescent protein,GFP）标记菌株PTS-GFP,构建其生长曲线,采用对峙生长法评价其室内抑菌活性,并应用抗生素平板回收结合激光扫描共聚焦显微镜观察标记菌株在番茄根围的定殖数量。结果显示：与原始菌株PTS-394相比,标记菌株PTS-GFP的生长、对青枯病菌和4种病原真菌的室内抑菌能力无明显差异。标记菌株PTS-GFP和青枯病菌菌液单独或混合处理番茄苗,灌根当天标记菌株初始菌量接近10⁸ CFU/g,处理3 d后种群数量迅速下降,约10⁶ CFU/g,随后缓慢下降,处理10 d后种群数量约10⁴ CFU/g,处理30 d后,标记菌株仍然能被检测到,约20 CFU/g。表明枯草芽胞杆菌PTS-394在番茄根际土壤中具有一定的定殖能力。     

拮抗内生细菌Bacillus mojavensisZA1在马铃薯根内及根际的定殖动态及其对土壤微生物的影响

利用GFP标记和平板分离法研究了拮抗内生细菌Bacillus mojavensisZA1在马铃薯根内及根际的定殖动态及其对土著微生物的影响。结果表明,标记菌株ZA1-gfp在土壤中定殖数量为3.7×10⁴~7.3×10⁴cfu/g;马铃薯根际为3.0×10³~1.5×10⁵cfu/g,根内为49~630cfu/g;马铃薯的根部横切面、根毛和根表皮等部位均观察到发绿色荧光的菌株ZA1-gfp菌体或聚集菌落。利用灌根法施入生防菌后,土壤中细菌、放射菌数量显著增加(P<0.05),土壤真菌数量在中期下降;土壤中细菌、真菌和放线菌的数量分别在相对稳定范围内(数据之间差异不显著)消长。该结果说明菌株ZA1-gfp能够在马铃薯根内及根际定殖且不影响土壤中土著微生物的群落稳定性。     

Arbuscular mycorrhizal fungal association in Asteraceae plants growing in the arid lands of Saudi Arabia

The present research was undertaken to explor the possibility of arbuscular mycorrhizal(AM) association with Asteraceae plants in the arid lands of Saudi Arabia(Al-Ghat, Buraydah, Thumamah and Huraymila). AM fungal colonization in the roots, spore numbers in the rhizosphere soil, fungal species diversity and correlation between AM properties and soil properties were determined. The highest colonization was in Conyza bonariensis(65%) from Al-Ghat, Anthemis cotula(52%) from Buraydah and C. bonariensis(53%) from Thumamah. The lowest was in Vernonia schimperi(41%) from Al-Ghat, Pulicaria undulata(25%) from Buraydah, Acanthospermum hispidum(34%) from Thumamah, Asteriscus graveolens(22%) and V. schimperi(22%) from Huraymila. Vesicular and arbuscular colonization were also presented in all plant species examined. The number of spores were 112–207 in Al-Ghat, 113–133 in Buraydah, 87–148 in Thumamah and 107–158 in Huraymila. Funneliformis mosseae, Glomus etunicatum, G. fasciculatum and G. aggregatum were identified. Relative frequency of AM fungal species varied widely and was irrespective of location and plant species. Diversity index varied with the rhizosphere soils of different plant species at various locations. Soil properties varied with locations and no distinct correlations were observed among the soil properties, root colonization and the number of spores. The results of the present study specified the association of AM fungi in different plants of Asteraceae and its significance in the ecological functioning of annual plants in the punitive environments of the rangelands in Saudi Arabia.     

果园杂草对枯草芽胞杆菌控制石榴枯萎病的增效作用

测定枯草芽胞杆菌(Bacillus subtilis)在石榴园6种优势杂草胜红蓟(Ageratum conyzoides)、三叶鬼针草(Bidens pilosa)、大籽蒿(Artemisia sieversiana)、鸭跖草(Commelina communi)、牛筋草(Eleusine indica)、牛膝菊(Galinsoga parviflora)根部及根际土壤中的定殖动态,及杂草与枯草芽胞杆菌协同控制石榴枯萎病(Ceratocystis imbriata)的效果,为生防细菌的应用提供新的方法.结果显示,枯草芽胞杆菌在胜红蓟、三叶鬼针草、大籽蒿和鸭跖草的根部定殖能力都相对较强.在大籽蒿的根部定殖能力最强.而枯草芽胞杆菌只有在大籽蒿根际土中的可检测浓度持续到接种后的30 d.各种杂草结合枯草芽胞杆菌防治石榴枯萎病的结果表明,大籽蒿和胜红蓟表现卓越的增效作用.表明石榴园部分优势杂草可能通过促进枯草芽胞杆菌在其根系及根际中的定殖从而增强控制石榴枯萎病的作用.     

Distribution and Biocontrol Potential of phlD(+) Pseudomonads in Corn and Soybean Fields

ABSTRACT The abundance and diversity of phlD(+) Pseudomonas spp. colonizing the rhizospheres of young, field-grown corn and soybean plants were assayed over a 3-year period. Populations of these bacteria were detected on the large majority of plants sampled in the state of Ohio, but colonization was greater on corn. Although significant variation in the incidence of rhizosphere colonization was observed from site to site and year to year on both crops, the magnitude of the variation was greatest for soybean. The D genotype was detected on plants collected from all 15 counties examined, and it represented the most abundant subpopulation on both crops. Additionally, six other genotypes (A, C, F, I, R, and S) were found to predominate in the rhizosphere of some plants. The most frequently observed of these were the A genotype and a newly discovered S genotype, both of which were found on corn and soybean roots obtained from multiple locations. Multiple isolates of the most abundant genotypes were recovered and characterized. The S genotype was found to be phylogenetically and phenotypically similar to the D genotype. In addition, the novel R genotype was found to be most similar to the A genotype. All of the isolates displayed significant capacities to inhibit the growth of an oomycete pathogen in vitro, but such phenotypes were highly dependent on media used. When tested against multiple oomycete pathogens isolated from soybean, the A genotype was significantly more inhibitory than the D genotype when incubated on 1/10x tryptic soy agar and 1/5x corn meal agar. Seed inoculation with different isolates of the A, D, and S genotypes indicated that significant root colonization, generally in excess of log 5 cells per gram of root, could be attained on both crops. Field trials of the A genotype isolate Wayne1R indicated the capacity of inoculant populations to supplement the activities of native populations so as to increase soybean stands and yields. The relevance of these findings to natural and augmentative biocontrol of root pathogens by these bacteria is discussed.